In the course of studying extracellular matrix synthesis by various childhood tumors, we have encounterd a previously unidentified, high molecular weight (500,000 D) protein secreted into the conditioned medium of several tumor cell lines. This protein is secreted in connection with laminin, type IV collagen, and fibronectin, but is immunologically distinct therefrom. It is also nonreactive with antibodies to basal lamina proteoglycan. It fails to label with radiolabeled sodium sulfate or glycosyl precursors, which further distinguishes it from conventional proteoglycans. Selective enzymatic degradation studies indicate that the protein is trypsin and pepsin sensitive, but collagenase and GAG degrading enzyme insensitive. Ultrastructural studies of rotary shadowed, purified molecules reveal a single, unbranched chain of over 700 nm length. The molecule co-purifies with laminin and type IV collagen under normal circumstances. Current efforts are aimed at devising preparative purification techniques which will allow purification of quantities sufficient to initiate the generation of monoclonal antibodies and biologic function studies.